I would like to detect cleaved caspases (17,19 kd) so Is 15% SDS PAGE ok for this?
running SDS PAGE 25mA for 1hr and western transfer conditions i use 100V for 1hr. and any suggestions during collecting sample because detection of caspases is tough i read.
Which kind of samples (matrix) ado you have?
For the SDS you can go up to 20% even, but I think the beste way is to try it directly.
25 mA as running time after primary sample migration is ok. For the time, refer to the marker bands.
For the western blotting maybe 40 minutes could be enough. But as I said before, optimizing the times it part of the method development. Indicatively, 40 minutes one hour should be enough.
ps: I think that the hard part about the caspase that you are talking about refers to some previous sample preparation step.
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